Patient samples used for single-cell RNA sequencing and whole exome sequencing.

Comparison to historical control comparing beta-binomial distributions.

Transarterial embolization in combination with photothermal therapy (PTT) represents a promising strategy for unresectable hepatocellular carcinoma (HCC). However, its clinical application is hindered by the lack of accurate laser-dosage guidance regimens, which results in tumor recurrence from insufficient irradiation or complications from excessive ablation. Here, we develop a depth-dependent laser-dosage guidance strategy for photothermal therapy via dual photoacoustic (PA)-Raman imaging using BBTPPRO-loaded poly(lactic-co-glycolic acid) microspheres (BBTPPRO MPs) as a theranostic embolic platform. The BBTPPRO MPs have a uniform spherical morphology (53.21 μm average diameter) with a drug loading of 4.73% and an encapsulation efficiency of 99.23%. The aggregated state of BBTPPRO through microsphere encapsulation offers substrate-free Raman signals via the stacking-induced intermolecular charge transfer-enhanced Raman scattering effect and enhances PA signals and photothermal conversion efficiency (29.7%) through the aggregation-caused quenching effect. By calibrating the required irradiation time to reach the temperature threshold required for complete thermal ablation under a fixed laser power density at various tissue depths, we establish a depth-dependent laser-dosage guidance strategy. In the orthotopic N1S1 rat liver tumor model after hepatic arterial embolization of BBTPPRO MPs, the PA imaging noninvasively assesses tumor depth to guide laser dosage, while the intraoperative Raman imaging provides tumor localization and delineation for the laser irradiation field. This dual-modal strategy has exhibited superior antitumor efficacy evidenced by a significantly lower maximum standardized uptake value via 18F-fluorodeoxyglucose positron emission tomography-computed tomography (1.68 ± 0.30) at 14 days post-treatment compared to conventional PTT group (6.92 ± 3.63). Overall, this PA-Raman image-guided strategy integrates pretreatment laser-dosage guidance with intraoperative spatial targeting, highlighting its potential to improve the safety and efficacy of HCC ablation.

Single-cell landscape of TME and epithelial cells from longitudinally collected tumor samples at baseline and during trial therapy.

Idiopathic pulmonary fibrosis (IPF) is a progressive and fatal interstitial lung disease with limited therapeutic options, highlighting the need for new molecular targets and effective antifibrotic agents. In this study, we investigated the role of B-cell lymphoma 9 (BCL9) in IPF and evaluated the antifibrotic potential of ZD-4009, a novel small-molecule inhibitor targeting BCL9. BCL9 was upregulated in fibrotic lung tissues, suggesting a possible association between BCL9 and pulmonary fibrosis progression. ZD-4009 showed high-affinity binding to β-catenin (pKD = 7.36 ± 0.01, KD = 44.72 nM). In vitro, ZD-4009 inhibited fibroblast activation, suppressed extracellular matrix deposition, and reduced the expression of fibrosis-associated markers, including COL1A1 and α-SMA. Mechanistically, ZD-4009 suppressed HIF-1α-associated glycolytic reprogramming and inhibited TGF-β/SMAD signaling, suggesting that its antifibrotic effects are associated with coordinated regulation of metabolic and profibrotic pathways. In vivo, ZD-4009 treatment was associated with alleviated bleomycin-induced pulmonary fibrosis, as reflected by improved survival from 55% in the bleomycin group to 82% in the bleomycin + ZD-4009 group, reduced relative collagen content from 61% to 48%, and ameliorated micro-CT-assessed fibrotic changes. In the tested model, ZD-4009 showed favorable antifibrotic effects compared with nintedanib, while liposome-encapsulated ZD-4009 reduced treatment-related toxicity. Together, these findings nominate ZD-4009 as a promising antifibrotic candidate and highlight BCL9 as a potential therapeutic target for IPF.

Hepatocellular carcinoma (HCC) is a malignancy characterised by high incidence and mortality rates, with China accounting for over 50% of new cases and deaths globally. The prevention and treatment of HCC remain formidable challenges, primarily due to pronounced tumour heterogeneity and a complex immune microenvironment. Recent advances in immunotherapy have highlighted the critical role of tumour antigen-specific T cells (TASCs) in the management of HCC. This review systematically examines the mechanisms underlying TASCs function, recent advances in HCC research, and the clinical significance of TASCs detection. It further analyses current challenges in HCC immunotherapy and offers a theoretical foundation and prospective directions for the development of combination and personalised therapies targeting TASCs.

3519 Background: Chemoimmunotherapy is standard frontline therapy for metastatic anal cancer, with no effective treatment options afterward. Mutation profiling has not yielded matched targeted therapies against anal cancer. Trophoblast cell-surface antigen 2 (TROP2) is overexpressed in other solid tumors, and TROP2 antibody-drug conjugates (ADCs) have demonstrated efficacy in breast and lung cancers. Given the therapeutic availability against TROP2, we evaluated expression of TROP2 in anal cancer as an actionable therapeutic target. Methods: Metastatic anal cancer tumors from 33 patients at MD Anderson were sequenced by bulk RNA sequencing (“Tumor Portrait” assay, Boston Gene) to quantify TACSTD2 (TROP2) gene expression. TACSTD2 expression was compared across all solid tumors and classified according to ranked expression percentile: high (>83 rd percentile), medium (17-83 rd percentile), or low (<17 th percentile). Gene expression (Nanostring) for >18,000 genes was measured using whole-genome digital spatial profiling (DSP) on a separate cohort of 40 chemoradiotherapy-refractory localized anal cancers collected at salvage surgery. TACSTD2 gene expression on tumor cells vs TME cells was compared with a t-test (SPSS). TROP2 protein expression (Invitrogen) was quantified by H-score using immunohistochemistry (IHC) and correlated with TACSTD2 expression via Spearman’s correlation. Three patient-derived xenograft (PDX) models of anal cancer (C1411, C1436, O0026) were treated with normal saline (untreated control, UTC) or the TROP2 ADC sacituzumab govitecan (sac-gov; 10 mg/kg IP twice weekly). Tumor growth inhibition (TGI) was defined as 1 - (mean tumor volumes of sac-gov/UTC) at 21 days. Results: Median TACSTD2 (TROP2) gene expression for patients with metastatic anal cancer was ranked at the 75 th percentile (IQR 63-85) relative to all solid tumors: 13/33 (39%) with high, 20/33 (61%) with medium, and none with low TACSTD2 expression. For localized anal cancers, TACSTD2 gene expression using DSP was significantly higher on tumor segments vs TME segments (log fold change 3.49, adjusted p < 0.0001). 30/39 (77%) anal cancers had high TROP2 expression by IHC, defined by H-score >200. Correlation between TACSTD2 gene expression and TROP2 protein expression by IHC was observed (r= 0.43, p < 0.001). In PDX models, TGI with sac-gov relative to UTC was observed in both models with high TROP2 expression (C1411: TGI 54%, p< 0.001; C1436: TGI 52%, p< 0.001) but not in a model with no TROP2 expression (O0026: TGI 12%, p=n.s.). Conclusions: TROP2 expression is high for localized and metastatic anal cancer. TACSTD2 gene expression correlates with matched TROP2 protein expression for anal cancer. Anti-tumor efficacy of TROP2 ADCs in vivo supports a forthcoming trial of sacituzumab tirumotecan for patients with treatment-refractory metastatic anal cancer, with the ultimate goal of establishing TROP2 as a predictive biomarker for treatment benefit in this rare cancer.

List of Patient Mosaic Team and Rare Tumors Initiative collaborators

Background/Objectives: Aberrant metabolism in tumors exacerbates the immunosuppressive tumor microenvironment. The immunosuppressive metabolite kynurenine inhibits the activation of effector T cells. Current antitumor drugs targeting kynurenine focus on small molecule inhibitors, which exhibit suboptimal efficacy in suppressing kynurenine generation owing to the diversity of kynurenine synthesis pathways. In contrast, kynureninase (KYNase) can directly metabolize kynurenine regardless of the production source. However, its delivery is hindered by short blood-circulation half-life and poor tumor accumulation. Additionally, photodynamic therapy (PDT) has been reported to synergize with immunotherapy, suggesting a potential combinatorial photodynamic immunometabolic cancer therapy with KYNase. Methods: A KYNase-Fc fusion protein was prepared to prolong blood circulation and enhance tumor accumulation of KYNase. Meanwhile, KYNase-Fc served as a nanocarrier for photosensitizer pheophorbide A (PhA) due to the high binding affinity between KYNase-Fc and PhA. Through self-assembly, KYNase-Fc/PhA nanoparticles (KYNase-Fc/PhA NPs) were prepared without extra carrier materials. Results: Compared with the PEGylated KYNase, KYNase-Fc exhibited significantly prolonged blood circulation, enhanced tumor accumulation and effective tumor suppression. Moreover, the prepared KYNase-Fc/PhA NPs facilitated rapid PhA tumor accumulation. The combined photodynamic immunometabolic therapy alleviated the immunosuppressive microenvironment and significantly inhibited the growth of subcutaneous 4T1 tumors in mice. Conclusions: KYNase-Fc offered a carrier-free nanomedicine for co-delivery of PhA for photodynamic immunometabolic antitumor therapy with enhanced efficacy, providing a promising platform for clinical translation.

Operating characteristics under varying trial-limiting toxicities rates.

Raman spectroscopy based on surface enhanced Raman scattering (SERS) has received great interest in biological and medical applications owing to its high specificity and sensitivity. However, a reliable and quantitative SERS analysis in vivo is a great challenge since SERS is affected by the unevenly distributed field enhancement in hot spots of nanoparticle substrate and unpredictable nanoparticle aggregation in the complex biological environment. Here, we present non-invasively quantitative Raman imaging through stacking-induced intermolecular charge transfer-enhanced Raman scattering (SICTERS) without relying on substrate. Based on the Raman imaging with SICTERS micelles, a quantitative method is established to accurately calculate the concentration of micelles in draining lymph nodes (DLNs) following subcutaneous administration. A physiologically based pharmacokinetic model is generated to fit the concentration-time curve of the SICTERS micelles in DLNs (R2 > 0.99), which calculates the pharmacokinetic parameters of the lymphatic transport of micelles with different sizes and surface charges. The optimized SICTERS micelles loading ovalbumin (OVA) as model antigen exhibits significant anti-tumor effect in mice bearing B16-OVA melanoma following vaccination. This study demonstrates a quantitative in vivo Raman imaging platform based on SICTERS for image-based pharmacokinetic modeling of micellar transport in lymphatics.

Background: To investigate the cause of ocular hypotension following retinal vein occlusion (RVO) and to elucidate the effect of vascular endothelial growth factor (VEGF) on intraocular pressure (IOP) reduction.Methods: A retrospective analysis was performed in 109 treatment-naïve unilateral central retinal vein occlusion (CRVO) patients to compare IOP between affected and fellow eyes. A subsequent prospective, cross-sectional study included 14 CRVO patients (before anti-VEGF therapy) and 27 cataract controls, from whom aqueous humor samples were acquired for VEGF measurement by ELISA. In animal experiments, rats received intravitreal VEGF injection in the right eye and vehicle in the left eye. IOP was measured at multiple time points post-injection. Trabecular meshwork tissues were collected at 4 hours for Western blot, PCR, and immunofluorescence analysis of Akt and eNOS expression.Findings: IOP was significantly lower in CRVO-affected eyes than in fellow eyes. The magnitude of IOP reduction correlated with aqueous VEGF levels. Additionally, eyes with presumed ischemic CRVO exhibited a significantly greater reduction in IOP and elevated aqueous VEGF levels compared with their non-ischemic counterparts. Subsequent animal experiments revealed that intravitreal injections of VEGF in rats significantly lowered IOP, an effect mediated by upregulation of the Akt/eNOS signaling pathway in the trabecular meshwork.Interpretation: VEGF reduces IOP via activation of the Akt/eNOS cascade in the conventional outflow pathway, contributing to RVO-associated ocular hypotension. Our findings suggest that the extent of IOP reduction may serve as a biomarker for RVO severity, and components of this pathway offer potential targets for novel glaucoma treatments.

Treatment emergent adverse events at least possibly related to study therapy.

Overall survival from treatment initiation with ixazomib plus gemcitabine plus doxorubicin.

Number of tumor cells in scRNA-seq analysis from the 11 patients with RMC.

This study reports the successful preparation of a highly active NiCo2O4/biochar composite catalyst (CNBC) and systematically investigates its performance in peroxymonosulfate (PMS)-activated degradation of 4-chlorophenol (4-CP). The biochar carbonized at 600 degrees C exhibits a defect-rich, partially graphitized structure effectively inhibits NiCo2O4 aggregation and promotes interfacial electron transfer, thereby significantly enhancing catalytic activity. At pH 6.7, with 0.3 g L-1 of CNBC and 0.7 mM PMS, the catalyst achieved complete 4-CP degradation within 60 min, a mineralization efficiency of 67.3%. The reaction tolerated coexisting substances well and remained stable after repeated use. Mechanism studies showed that the reaction mainly followed a non-radical route. Singlet oxygen and direct electron movement through a short-lived CNBC-PMS* activated state played key roles. Surface groups such as C--O and-OH, together with adsorbed oxygen, acted as the main reaction centers. In addition, the reversible change between Co and Ni oxidation states helped sustain the catalytic activity. Liquid chromatography-mass spectrometry analysis and toxicity assessments indicated that the nine detected intermediates posed significantly lower ecological risks. Overall, the CNBC/PMS system exhibited excellent degradation efficiency, structural stability, and environmental compatibility. These findings provide both a material basis and theoretical support for the green oxidative remediation of chlorinated aromatic pollutants.

Low-grade serous carcinoma of the ovary (LGSOC) is relatively resistant to chemotherapy. Given its biological parallels to hormone receptor-positive breast cancer, including responsiveness to anti-estrogen therapies, we conducted a pilot phase II study to assess clinical benefit of neoadjuvant fulvestrant and abemaciclib in women with advanced, unresectable LGSOC (NCT03531645). Imaging assessments were performed every 8 weeks until resectable. The primary endpoint was clinical benefit rate (CBR). Exploratory objectives included evaluation of safety profile, accessing biomarkers related to clinical benefit, and description of tumor phenotypic changes. Fifteen patients were enrolled and evaluable for efficacy. CBR was 100%, with 1/15 patients (7%) achieving complete response (CR), 8/15 patients (53%) achieving partial response (PR), and 6/15 (40%) exhibiting stable disease (SD). Interval cytoreductive surgery (ICS) was performed in 9 patients (60%), with 7 (78%) achieving complete or optimal resection. Fourteen tumor samples underwent transcriptomic and proteomic profiling. Tumors from long-term survivors showed significantly higher baseline expression of cell-cycle-related programs and estrogen signaling-related genes, both suppressed upon treatment. Neoadjuvant fulvestrant and abemaciclib was well tolerated, achieved high response rates, and enabled optimal surgical resection in most patients. Tumor proliferative activity and estrogen signaling dependency may predict therapy response.